Method of preparing zein solutions directly from gluten



Patented Sept. 26, 1944 METHOD OF PREPARING ZEIN SOLUTIONS;

' DIRECTLY FROM GLUTEN Roy E. Golemam Chicago, 1111., assignor to Time, incorporated, a. corporation of New York 9 No Drawing. Original application November 30,

1942, Serial N0. 167,4412. Divided and this application October 25, 1943, Serial, No. 507,596

10 Claims.

This invention relates to methods of preparing commercially usable solutions and coating compositions of p'rolamines or prolamine-containing proteins directly from glutens, either corn, wheat, barley, etc. methods which will produce solutionsand coating compositions oi? the alcohol-soluble portion of corn gluten, of controlled solution and gelling characteristics.

. This applicationis a division of my copending application, Serial No. 467,402, filed November 30, 1942.

'Heretofore, in the preparation of zein solutions and coating compositions, including the substantially non-aqueous zein solutions and coating compositions of controlled solution and gelling characteristics as described in my prior Patent No.'2,185,110, granted December 26, 1939, for example, the zein is initially isolated from corn gluten and then dissolved in suitable solvents to form the desired solutions. To isolate zein from It .is particularly directed to such preferably between about 140 to about 150 F., for a short period of time generally not exceeding about three hours. It is preferred'that the extraction time be not longer than about onehalf to about one hour since within this time and at the temperatures stated, the zein in the gluten is extracted and is not deleteriously efiected.

The extract solution obtained as above described is now separated from the residual gluten by filtration, pressing, centrifugal separation or in any other desirable manner and is distilled or otherwise treated to remove all or some of the 'ily to the step of removing the diluent, if necessary. The resulting residue isa. solution of zein or zein-containing protein in the base solvent corn gluten as acommercially usable powder is a I laborious and expensive process, involving steps of precipitation, settling, filtering, washing and drying. The solubility of the zein in zein solvents and the solution characteristics such as stability and gelling tendencies are largely dependent upon the care exercised in carrying out the steps of theprocess.

In accordance with the present invention I prepare solutions of zein or zein-containing proteins directly from corn gluten under conditions that will readily produce a wide variety of zein products in so far as properties and solubility characteristics are concerned, and thereby avoid the expense and objections incident to thepreliminary isolation of zein in powder form. In-

carrying out the present invention, corn gluten, for example, in granular'or powdered form, is treated witha suitable solvent mixture, without the establishment of special conditions of hydrogen ion concentration either in the solventor in the gluten, to extract the zein from the gluten.

The solvent mixture comprises a base solvent having a boiling point above about 125 0., preferably above about 150 C. and a diluent or extender having'a boiling P int below about 125 C. and, preferably, of about 100 C. and below.

The base solvent is itself a solvent for zein and the diluent may be, and preferably is, also a solvent for zein. The base solvents and diluents in accordance with the present invention will be defined fully hereafter.

The extraction is carried'ont at elevated temperatures, say from about 120 to about 170 F.,

iorm at the re-heating temperatures.

do not separate even when cooled to temperatures of 50 to F. and somewhat below. When cooled even to temperatures as low as 10 above zero or to zer0,,solutions prepared in accordance with the present invention may separate and become solid; however, on heating again to temperatures of from about 45 to 70 E, a re-solution is efiected, either without stirring or with slight stirring, and the solutions return to their. normal eral, these solutions are substantially non-gelling and have but a slight tendency to. increase in viscosity; that is, to thicken or become heavier in body. In many instances they exhibit practically no tendency to thicken or to gel over practical periods of time in the order of about 3 to 6 months and even up to one year and longer. These solutions remain stable and reasonably constant in their characteristics during the period necessary for transportation and distribution, either in bulk or in packages, or for storage for reasonable periods of time, and hence their field of applicability is greatly increased.

In general, in the concentration of zein or other prolamine in commercially usable solutions containing about 12 to 30% of protein, the

amount of base solvent necessary to obtain these In genconcentrations in the final solutions is in and of itself insufficient to give the proper relation between solvent and gluten for effective extraction results. If large amounts of the relatively high boiling point base solvents are used, sumoient to give the proper relation between solventand gluten, the extraction results are not sumciently effective and, moreover, the cost of treating the extracts to obtain the desired solids concentration is so prohibitive as to make this procedure commercially unfeasible.

The desired solvent to gluten ration in accordance with the present invention is secured by adding to the desired amount of relatively high boiling point base solvent or mixture of these solvents one or more of the so-called relatively low boiling point diluent solvents. To obtain efiective extraction results, the solvent to gluten ratio should be in the order of about 2 to parts by weight of the former to about 1 part by weight of the latter. Due to the presence of a diluent or extender in the extracting solvent, the solvent to gluten ratio may .be less than that which would be required with a base solvent alone and, notwithstanding the lower solvent to gluten ratio, the extraction results areggenerally more efiective. have a lower boiling point and distillation range than the base solvent. It is preferred that these diluent solvents or extenders be of substantially lower boiling point and distillation range than the base solvent, as pointed out above.

Base solvents The base solvents are relatively high boiling compounds or mixtures of compounds as pointed out above and have a suitable balance between the hydrocarbon constituents or radicals present in the solvent or solvent mixtures and the polar constituents or radicals present therein. A certain proportion of hydroxyl radicals in the base solvents is required to secure solvent action. Other polar radicals such as --0 (ether oxygen), -Cl, NH2 and COOH appear to aid or cooperate with the OH in securing the necessary conditions of polarity in the base solvents (in which term I include mixtures of base solvents) for solvent action.

The proportion of polar'radicals to hydrocarbon or non-polar radicals which I have found to be desirable lies within a range which extends between the limits of absolute methanol on the one hand, having 53% hydroxyl and 47% hydrocarbon radicals, and absolute ethanol on. the other hand having 37% hydroxyl and 63% hydrocarbon radicals. The benzy1 radical acts as if it were intermediate the methyl and ethyl radicals. This range is approximate and may vary somewhat depending on the solvent or solvent mixture used. The limits of this range are indicative of solvent properties and provide a close and adequate guide to enable the operator, by simple experimentation, to determine the required balance of polar and hydrocarbon radicals in the base solvent or mixture of base solvents in accordance with the present invention.

It may be stated that the base solvents in accordance with the present invention may be any one ofthe solvents or mixture of solvents described in my prior Patent No. 2,185,110 which has the required balance of polar and non-polar radicals, and a boiling point above about 125 0., preferably, above about 150 C. as pointed out The diluent solvent or extender must 1 the present invention are the glycols such as-di-' ethylene glycol, propylene glycol, triethylene glycol and mixtures thereof, including mixtures which contain ethylene glycol; diacetone alcohol; closed chain cyclic alcohols such as benzyl alcohol, cyclohexanol, fu'rfuryl alcohol, tetrahydrofurfuryl alcohol and mixtures of closed chain alcohols; glycol ethers such as ethylene glycol mono-methyl ether, diethylene glycol monomethyl ether, ethylene glycol mono-ethyl ether, diethylene glycol monoethyl and mixtures of glycol ethers; mixtures of two or more of the foregoing enumerated base solvents; mixtures of one or more of the foregoing base solvents with glycerine, or other solvents or mixtures of solvents, providing the mixture has the required polar to non-polar radical balance and the required relatively high boiling point. It is to be understood that the foregoing specifically mentioned solvents or mixtures of solvents is not intended to be inclusive of all the base solvents which may be used in accordance with the present invention since they are merely set forth for illustrative purposes. Y

- Diluent solvents The diluent solvents or extenders in accordance with the present invention may be, suitably, a single solvent or a solvent mixture may or may not be a solvent for zein or other prolamine, as

' hydrous methanol or in excess of that present above. For a more complete description of the kinds of solvents which mav be used as base solin the azeotropes alcohol and 91% isopropyl alcohol. Greater or lesser amounts of water than as stated may be used; however, best results are obtained with the use of added amounts of water within the percentage range stated. For convenience herein I term these alcoholic solvents aqueous alcohol mixtures.

Another class of diluent solvents in accordance with the present invention comprises mixtures of an azeotropic concentrated alcohol solvent with a non-solvent and water. Examples of such diluents are mixtures of 95% ethyl alcohol or 91% isopropyl alcohol with dichlorethylene, trichlorethylene, ethyl acetate, benzene, cyclohexane, etc., and water. In this class of diluent solvents- I also include non-aqueous azeotropic mixtures of anhydrous methyl alcohol with hexane, cyclohexane, benzene, carbon tetrachloride, dichlorethylene, trichlorethylene, acetone, methyl acetate, ethylacetate, methyl ethyl ketone, etc, because one constituent of the mixture, methyl alcohol, is a zein solvent.

In addition to .the foregoing, wherein at least one constituent of a diluent solvent mixture is a.

solvents wherein the individual constituents of the mixture are non-solvents for zein proteins.

'Water. azeotropic mixtures of anhydrous ethyl alcohol stant boiling point mixture containing 80% dioxan boiling at 86.9 C. and butyl alcohol forms with water a constant boiling point mixture containing 63% butyl alcohol boiling at 92 C. They may also be suitable aqueous azeotropic mixtures which are non-solvents for zein proteins such a ethyl methyl-'ketone' and And they may also be non-aqueous or anhydrous isopropyl alcohol with hexane, cyclohexane, benzene, carbon tetrachloride, dichlorethylene, trichlorethylene, acetone, methyl acetate, ethyl acetate, methyl ethyl ketone, etc. In general, extracting solvents consisting of or including the aqueous alcohol mixtures are capable of extracting larger amounts of zein protein from gluten than are the extracting solvents consisting of or including a concentrated alcohol solvent. It has been noted that for, practical extraction procedures, using either a concentrated alcohol solvent alone or an aqueous alcohol mixture alone, approximately 2 parts by weight of solvent to 1 of gluten may be used. At this concentration, with gluten containing approximately 60% total protein, the extract solutions of the concentrated alcohol sol'vent con-=1 tain a maximum'zein protein content of approximately whereas the extract solutions.

of the aqueous alcohol mixture solvents contain a maximum zein protein content of from about 12% to about 15% and even up to In addition,-the aqueous alcohol mixture solvents are capable of extracting a substantially larger pro-' portion of a desirable type of zein present in the protein of the gluten than alcohol solvents.

Zein solutions of a desired concentration and of a'wide applicability can be obtained directly from gluten by having in the final solution as the only solvent or asthe main solvent for zein, a relatively high boiling point solvent having the required balance of polar and non-polar radicals as described herein. It is now well established are the concentrateda that the viscosity, gelling tendencies, and prac-,

ticalusability of zein solutions are in no small part dependent upon the character of the solvent in which the zein is dissolved.

'Zein or zein-containing extract solutions containing. any one or more of the foregoing base solvents inaccordance with the present invention, may be boiled, distilled or otherwise treated toremove part or all of the diluent. It is'evldent that practically'all of the diluent, including any water carried from the gluten to the extract solution, may be removed, thereby obtaining solutions of zeinor zein-containing proteins,'in the base solvent which, for all practical punposes, are substantially anhydrous.

Such solutions vary widely in character, depend- 'ing upon the choice of 'solvents or solvent mixtures used, as is to be expected, and some of them may undergo changes (apparent insolubility, for example), during the diluent and water removal which affect the solvent characteristics of thezein and zein-containing proteins in the base solvent utilized. These changes,

where they take place, are not of a permanent other highly hydroxylated compound, such as methyl or ethyl alcohol or a mixture of such compounds.

In the preparations of usable solutions having amounts stated. While'water is, in general,

not an essential constituent of some of the solutions formed in accordance with the present invention, the solutions are generally of an improved character when. they contain small amounts of water. The solutions may contain larger amounts of water, say between 5 to 10%, or even more.

As already pointed out, the type or character of thezein-containing protein produced by the extraction is in no small part dependent on the character of the extraction solvents used. This condition'is particularly pronounced when the highly aqueous alcohols or the high water azeotropes such as dioxan-water or n-butyl alcohol.

7 water mixtures are used as diluents. Not only do these diluents efiect the extraction ofa larger characterand it is possible to effect a re-solution of the zein or zein-containing proteins in the base solvents by the addition of a small amount of water, say'in the order of from about 1 to 3% by weight, or an equivalent amountof amount of the zein-containing protein than do the more concentrated alcohols, but they also produce by their extraction a. type of zein protein which generally produces solutions having a high viscosity and pronounced colloidal consistency. On the other hand the zein protein produced with the concentrated alcohols and less aqueous diluents tends to produce solutions having a lower viscosity and a lesser colloidal consistency. It isthus evident that various types of zein protein having substantially diflerent properties can be produced by proper selection and composition of the diluents.

From the foregoing it is manifest that the natureand properties of the finished solution will be determined by the nature o f ihelsase solvent, the amount of zein protein presentin the solution and the type or character'ot "the zein protein extracted by the particular diluent used. It

is apparent that with the solvent and diluent combinations possible, a wide variety of solutions for many uses can be produced by the methods of the present invention.

Withinthe range of the many possible combinations and the methods in accordance with the present invention, solutions may be produced which require further adjustment or modification in order to secure improved solution or desired stability and controlled gelling properties. Such adjustments or modifications can be readily effected bythe use of rosin, fatty acids, compatible amines such as monoethanolamine, triethanol- .amine, etc., amine soaps of rosin or fatty acids, or by the various methods and materials for producing stable, controlled gelling, zein solutions described in my issued Patents Nos. 2,185,122, 2,246,779 and 2,298,548. The addition of suitable materialsfor effecting desired modification or adjustment may be made to the extract solution before distilling oi the diluent or to the final solution. Tire preferable procedure and materials with particular solutions may be easily determined by experimentation.

In the following examples I disclose illustrative method for producing zein or zein-containing solutions directly from gluten in accordance with the present invention. In the examples and glycol was added to 30 parts of powdered gluten in a vessel. The mixture was heated to about 160 F.'and held atabout that temperature for 7 about /2- hour with stirring to extract the zeincontaining proteins from the gluten. The mixture was then forced through a filter to separate the zein-containing extract and the extract was then slowly heated to about 250 F. and held at about that temperature with stirring until cessation of visible boiling. The residue was a solution of zein-containing proteins principally in the mixture of glycols.

In the following examples the manipulative procedures were the same as those set forth in Example 1, with the exceptions hereinafter noted, and in each instance the extracts were heated until cessation of visible boiling.

Example 2.-A solvent mixture containing 72 parts 'of a mixture consisting of 80% of dioxan and of water, and 48 parts of diethylene glycol was added to parts of powdered gluten. The procedure .was in Example 1 and the resulting residue was a solution of zein-containing proteins principally in diethylene glycol.

Example 3.--A solvent mixture containing 60 parts of a mixture consisting of 63% of n-butanol and 37% of water, and 40 parts of diethylene glycol was added to 30 parts of powdered gluten. The procedure was as'in Example 1 except that the extraction temperature was 140 F. The resulting residue was a solution of zein-containing proteins principally in diethylene glycol.

Example 4.-A solvent mixture containing 60 parts of a mixture consisting of 88.4% of ethyl methyl ketone and 11.6% of water, and 40 parts of diethylene glycol was added to 30 parts of powdered gluten. The procedure was as in Example 1 except that the extraction temperature was 140 F. The resulting residue was a solution of zein-containing proteins principally in diethylene glycol.

At the cessation of boiling as referred to in the examples some proportion of the lower boiling constituents of the diluent originally used remains in the final solution. thereof so remaining depends upon the final temperature and the boiling point curve of the mix ture. Considerable proportions of the diluent or of its constituents, including water, may remain without causing gelling of the final solution in The proportion by these zein-containing solutions and coating compositions are very strong and have adherent properties. In these solutions and coating compositions the film forming properties thereof can tion, Serial No. 467,402.

While my invention has been described in conv nection with certain specific examples, it is, of

reasonable periods of time. This is particularly true where the glycols are used. In this respect the final solutions show differences from solutions of r commercial zein containing large amounts of water as in the prior art, apparently by reason of some changes in the character or structure of the zein-containing proteins of the corn gluten taking place in the process of the present invention, the nature of which changes is not as yet understood.

In general and influenced by the type and amount of zein protein present, the solutions prepared as described above and the coating compositions prepared therefrom have the properties, particularly when heated, of.being able to give up their solvent or solvents. readily, and when applied to any surface they quickly and readily form tough, flexible, non-tacky, hard and generally transparent coatings even when retaining some solvent. The coatings formed course, obvious it is not to be construed as limited to these examples or to the details of the methods set forth therein, since obvious changes in materials, proportions and method details will be apparent'from the foregoing.

' In the claims, the expression an elevated temperature in the order of about F. to about 170 F. is not to be construed as a precise critical range, but, rather, as a range of temperatures indicative of the elevated temperature at which the extractions may be carried out.

I claim:

1. The method of preparing directly from gluten a. solution of zein-containing proteins in a solvent or mixture of solvents comprising as an essential solvent constituent for the zeincontaining proteins, a solvent havin a boiling point above about C. and having a proportion of polar to non-polar radicals which lies within a range extending between the limits of the proportion of polar to non-polar radicals of methanol and the proportion of polar to nonpolar radicals of ethanol, which comprises contacting gluten with a solvent mixture at an elevated temperature in the order of about 120 F. to about 170 F. to extract zein-containing proteins from the gluten, said solvent mixture comprising the aforesaid zein-containing protein solvent havinga boiling point above about 125 C. and a miscible diluent therefor having a boiling point below about 125 C., said diluent being an azeotropic mixture which is a solvent for zein wherein the constituents of the mixture are individually non-solvents forzein, separating the extract from the residual gluten and removing point above about C. and having a propor- 1 tion of polar to non-polar radicals which lies within a range extending between the limits of the proportion of polar to non-polar radicals of methanol and the proportion of polar to nonpolar radicals of ethanol, which comprises contacting gluten with a solvent mixture at an elevated temperature in the order of about 120 F. to about F. to extract zein-containing proteins from the gluten, said solvent mixture comprising the aforesaid zein-containing protein solvent having a boiling point above about 150 C. and a miscible diluent therefor having a boiling point below about 12550., said diluent being an azeotropic mixture which is a solvent for zein wherein the constituents of the mixture are individually non-solvents for zein, separating the extract from the residual gluten and removing substantially all of the diluent from the extract to form the aforesaid solution.

3. The method of preparing directly from gluten a solution of zein-containing proteins in a solvent or mixture of solvents comprising as an essential solvent constituent for the mincontaining proteins, a solvent having a boiling point above about 150 C. and having a proportion of polar to nonlar radicals which lies within a range extending between the limits of th proportion of polar to non-polar radicals of methanol and the proportion of polar to nonpolar radicals of ethanol, which comprises contacting gluten with a solvent mixture at an elevated temperature in the order of about 120 F. to about 170 F. to extract zein-containing proteins from the gluten, said solvent mixture comprising the aforesaid zein-contalning protein solvent having a boiling point above about 150 C. and a miscible diluent therefor having a boiling point below about 125 C., said diluent bein an azeotropic mixture which is a. solvent for zein wherein the constituents of the mixture are individually non-solvents for zein, separating the extract from the residual gluten and removing at least part of the diluent from the extract to' form the aforesaid solution.

4. The method of preparing directly from gluten a solution of zein-containing proteins in a solvent or mixture of solvents comprising a glycol as'an essential solvent constituent for the zein-containing proteins, which comprises contacting gluten with a solvent mixture at an elevated temperature in the order of about 120 F. to about 170 F. to extract zein-containing pro teins from the gluten, said solvent mixture comprising a glycol and a miscible diluent therefor having a boiling point below about 125 C., said diluent being an azeotropic mixture which is a solvent for zein wherein the constituents of the mixture are'individually non-solvents for zein, separating the extract from the residual gluten and removing at least part of the diluent from the extract to form the aforesaid solution 5. The method of preparing directly from gluten a solution of zein-containing proteins in a solvent or mixture of solvents comprising a glycol as an essential solvent constituent for the zein-containing proteins, which comprises contacting gluten with a solvent mixture at an elevated temperature in the order of about 120 F.

to about 170 F. to extract zein-containing proteins from the gluten, said solvent mixture comprising a glycol and a miscible diluent therefor having a boiling point below about 125 0., said diluent comprising an azeotropic mixture of dioxan and water which is a solvent for said rein-containing proteins, separating the extract from the residual gluten and removing at least part of the diluent from the extract to form the aforesaid solution.

6. The method of preparing directly from gluten a solution of zein-containing proteins in a solvent or mixture of solvents comprising a gLvcol as an essential solvent constituent for the zein-containing proteins, which comprises contacting gluten with a solvent mixture at an ele vated temperature in the order of about 120' F. to about 170 F. to extract min-containing prosolvent or mixture of solvents comprising a gly-' teins from the gluten, said solvent mixture comprisin a glycol and a miscible diluent therefor having a boiling point below about 126 C., said diluent being a solvent for said zein-containing proteins-and comprises an azeotropic mixture of n-butyl alcohol and water, separating the extract from the residual gluten and removing at least part of the diluent from the extract to form the aforesaid solution.

7.- The method of preparing directly from gluten 9. solution of zein-containing proteins in a col as an essential solvent constituent for the zein-containing proteins, which comprises contacting gluten with a solvent mixture at an elevated temperature in the order of-about 120 F. to about 170 F. to extract zein-containing proteins from the gluten, said solvent mixture comprising a, glycol and a miscible diluent therefor having a boiling point below about 125 C., said diluent comprising an azeotropic mixture of ethyl methyl ketone and water which is a solvent for said zein-containing proteins, separating the extract from the residual gluten and removing at least part of the diluent from the extract to form the aforesaid solution.

8. The method of preparing directly from gluten a solution of zein-containing proteins in a solvent or mixture of solvents comprising diethylene glycol as an essential solvent constituent for the zein-containing proteins, which comprises contacting gluten with a solvent mixture at an elevated temperature in the order of about F. to about F.- to extract zein-contalning proteins from the gluten, said solvent mixture comprising diethylene glycol and an azeotropic mixture of dioxan and water, separating the extract from the residual gluten and removing at least part of the said azeotropic mixture from the extract to form the aforesaid solution.

9. The method of preparing directly from gluten a solution of zein-containing proteins in a solvent or mixture of solvents comprising diethylene glycol as an essential solvent constituent for the zein-containing proteins, which comprises contacting gluten with a solvent mixture at an elevated temperature in the order of about 120 F. to about 170 F. to extract zein-containing proteins from {the gluten, said solvent mixture comprising diethylen glycol and an azeotropic mixture of nbutyl alcohol and water, separating the extract from the residual gluten and removing at least part of the said azeotropic mixture from the extract to form the aforesaid solution.

10. The method of preparing directly from gluten a. solution of zein-containing proteins in a solvent or mixture of solvents comprising diethylene glycol as an essential solvent constituent for the zein-containing proteins, which comprises contacting gluten with a solvent mixture at an elevated temperature in the order of about 120 F. to about 170 F. to extract zein-containing proteins from the gluten, said solvent mixture comprising diethylene glycol and an azeotropic mixture of ethyl methyl ketone and water, separating the extract from the residual gluten and removing at least part ture from the extract tion.

to form the aforesaid solu- ROY E. COLEMAN.

oi the said azeotropic mix- Patent No. 2, 9,202.

CERTIFICATE OF CORRECTION.

September 26, 19141;. ROY E. COLEMAN.

It is hereby certified that error appears in the printed specification of the above numbered patent requiring correction as follows: Page 2, first column, line 15, for the-word "these" read --base--; and second column,

line 55, after "inixflmre" insert "which"; line 56, for "extended" read "extender"; page Lg, first column, line 2?, after "was" insert --as--;

and that the said Letters Patent shoizld be read with this correction therein that the same may conform to the record of the case in the Patent 01- fice.

Signed and sealed this 5th da of December, A. D. 19%.

Leslie Frazer (Seal) Acting Commissioner of Patents. 

